In vivo MR imaging of intravascularly injected magnetically labeled mesenchymal stem cells in rat kidney and liver

PURPOSE: To evaluate in vivo magnetic resonance (MR) imaging with a conventional 1.5-T system for depiction and tracking of intravascularly injected superparamagnetic iron oxide (SPIO)-labeled mesenchymal stem cells (MSCs). MATERIALS AND METHODS: This study was conducted in accordance with French law governing animal research and met guidelines for animal care and use. Rat MSCs were labeled with SPIO and transfection agent. Relaxation rates at 1.5 T, cell viability, proliferation, differentiation capacity, and labeling stability were assessed in vitro as a function of SPIO concentration. MSCs were injected into renal arteries of healthy rats (labeled cells in four, unlabeled cells in two) and portal veins of rats treated with carbon tetrachloride to induce centrolobular liver necrosis (labeled cells and unlabeled cells in two each). Follow-up serial T2*-weighted gradient-echo MR imaging and R2* mapping were performed. MR imaging findings were compared histologically. RESULTS: SPIO labeling caused a strong R2* effect that increased linearly with iron dose; R2* increase for cells labeled for 48 hours with 50 microg of iron per milliliter was 50 sec(-1) per million cells per milliliter. R2* was proportional to iron load of cells. SPIO labeling did not affect cell viability (P > .27). Labeled cells were able to differentiate into adipocytes and osteocytes. Proliferation was substantially limited for MSCs labeled with 100 microg Fe/mL or greater. Label half-life was longer than 11 days. In normal kidneys, labeled MSCs caused signal intensity loss in renal cortex. After labeled MSC injection, diseased liver had diffuse granular appearance. Cells were detected for up to 7 days in kidney and 12 days in liver. Signal intensity loss and fading over time were confirmed with serial R2* mapping. At histologic analysis, signal intensity loss correlated with iron-loaded cells, primarily in renal glomeruli and hepatic sinusoids; immunohistochemical analysis results confirmed these cells were MSCs. CONCLUSION: MR imaging can aid in monitoring of intravascularly administered SPIO-labeled MSCs in vivo in kidney and liver.     

Clinically applicable labeling of mammalian and stem cells by combining superparamagnetic iron oxides and transfection agents

PURPOSE: To label mammalian and stem cells by combining commercially available transfection agents (TAs) with superparamagnetic iron oxide (SPIO) magnetic resonance (MR) imaging contrast agents. MATERIALS AND METHODS: Three TAs were incubated with ferumoxides and MION-46L in cell culture medium at various concentrations. Human mesenchymal stem cells, mouse lymphocytes, rat oligodendrocyte progenitor CG-4 cells, and human cervical carcinoma cells were incubated 2-48 hours with 25 microg of iron per milliliter of combined TAs and SPIO. Cellular labeling was evaluated with T2 relaxometry, MR imaging of labeled cell suspensions, and Prussian blue staining for iron assessment. Proliferation and viability of mesenchymal stem cells and human cervical carcinoma cells labeled with a combination of TAs and ferumoxides were evaluated. RESULTS: When ferumoxides-TA or MION-46L-TA was used, intracytoplasmic particles stained with Prussian blue stain were detected for all cell lines with a labeling efficiency of nearly 100%. Limited or no uptake was observed for cells incubated with ferumoxides or MION-46L alone. For TA-SPIO-labeled cells, MR images and relaxometry findings showed a 50%-90% decrease in signal intensity and a more than 40-fold increase in T2s. Cell viability varied from 103.7% +/- 9 to 123.0% +/- 9 compared with control cell viability at 9 days, and cell proliferation was not affected by endosomal incorporation of SPIO nanoparticles. Iron concentrations varied with ferumoxides-TA combinations and cells with a maximum of 30.1 pg +/- 3.7 of iron per cell for labeled mesenchymal stem cells. CONCLUSION: Magnetic labeling of mammalian cells with use of ferumoxides and TAs is possible and may enable cellular MR imaging and tracking in experimental and clinical settings.     

Capacity of human monocytes to phagocytose approved iron oxide MR contrast agents in vitro

To evaluate the capacity of human monocytes to phagocytose various approved iron oxide based magnetic resonance (MR) contrast agents and to optimize in vitro labeling of these cells. Human monocytes were incubated with two superparamagnetic iron oxide particles (SPIO) as well as two ultrasmall SPIO (USPIO) at varying iron oxide concentrations and incubation times. Iron uptake in monocytes was proven by histology, quantified by atomic emission absorption spectrometry and depicted with T2* weighted fast field echo (FFE) MR images at 1.5 T. Additionally, induction of apoptosis in iron oxide labeled monocytes was determined by YO-PRO-1 staining. Cellular iron uptake was significantly (P<0.01) higher after incubation with SPIO compared with USPIO. For SPIO, the iron oxide uptake was significantly (P<0.01) higher after incubation with the ionic Ferucarbotran as compared with the non-ionic Ferumoxides. Efficient cell labeling was achieved after incubation with Ferucarbotran at concentrations 500 g Fe/ml and incubation times 1 h, resulting in a maximal iron oxide uptake of up to 50 pg Fe/cell without impairment of cell viability. In vitro labeling of human monocytes for MR imaging is most effectively obtained with the approved SPIO Ferucarbotran. Potential subsequent in vivo cell tracking applications comprise, e.g. specific targeting of inflammatory processes.     

In vivo serial evaluation of superparamagnetic iron‐oxide labeled stem cells by off‐resonance positive contrast

MRI is emerging as a diagnostic modality to track iron‐oxide‐labeled stem cells. This study investigates whether an off‐resonance (OR) pulse sequence designed to generate positive contrast at 1.5T can assess the location, quantity, and viability of delivered stem cells in vivo. Using mouse embryonic stem cell transfected with luciferase reporter gene (luc‐mESC), multimodality validation of OR signal was conducted to determine whether engraftment parameters of superparamagnetic iron‐oxide labeled luc‐mESC (SPIO‐luc‐mESC) could be determined after cell transplantation into the mouse hindlimb. A significant increase in signal‐ and contrast‐to‐noise of the SPIO‐luc‐mESC was achieved with the OR technique when compared to a gradient recalled echo (GRE) sequence. A significant correlation between the quantity of SPIO‐luc‐mESC and OR signal was observed immediately after transplantation (R² = 0.74, P < 0.05). The assessment of transplanted cell viability by bioluminescence imaging (BLI) showed a significant increase of luciferase activities by day 16, while the MRI signal showed no difference. No significant correlation between BLI and MRI signals of cell viability was observed. In conclusion, using an OR sequence the precise localization and quantitation of SPIO‐labeled stem cells in both space and time were possible. However, the OR sequence did not allow evaluation of cell viability. Magn Reson Med 60:1269–1275, 2008. © 2008 Wiley‐Liss, Inc.     

Transferrin receptor upregulation: in vitro labeling of rat mesenchymal stem cells with superparamagnetic iron oxide

PURPOSE: To prospectively evaluate the influence of superparamagnetic iron oxide (SPIO) or ultrasmall SPIO (USPIO) particles on the surface epitope pattern of adult mesenchymal stem cells (MSCs) by regulating the expression of transferrin receptor and to prospectively evaluate the influence of transfection agents (TAs) on the uptake of SPIO or USPIO particles in MSCs. MATERIALS AND METHODS: The study was approved by the institutional animal care committee of the University of Tübingen. MSCs were isolated from the bone marrow of four rats. To obtain highly homogeneous MSC populations, MSCs from one rat were single-cell cloned. One MSC clone was characterized and selected for the labeling experiments. The MSCs, which were characterized with flow cytometry and in vitro differentiation, were labeled with 200 microg/mL SPIO or USPIO or with 60 microg/mL SPIO or USPIO in combination with TAs. Aggregations of labeled cells were accommodated inside a defined volume in an agar gel matrix. Magnetic resonance (MR) imaging was performed to measure SPIO- or USPIO-induced signal voids. Quantification of cellular total iron load (TIL) (intracellular iron plus iron coating the cellular surface), determination of cellular viability, and electron microscopy were also performed. RESULTS: Labeling of MSCs with SPIO or USPIO was feasible without affecting cell viability (91.1%-94.7%) or differentiation potential. For MR imaging, SPIO plus a TA was most effective, depicting 5000 cells with an average TIL of 76.5 pg per cell. SPIO or USPIO particles in combination with TAs coated the cellular surface but were not incorporated into cells. In nontransfected cells, SPIO or USPIO was taken up. MSCs labeled with SPIO or USPIO but without a TA showed enhanced expression of transferrin receptor, in contrary to both MSCs labeled with SPIO or USPIO and a TA and control cells. CONCLUSION: SPIO or USPIO labeling without TAs has an influence on gene expression of MSCs upregulating transferrin receptor. Furthermore, SPIO labeling with a TA will coat the cellular surface.     

Cell motility of neural stem cells is reduced after SPIO‐labeling,which is mitigated after exocytosis

MRI is used for tracking of superparamagnetic iron oxide (SPIO)‐labeled neural stem cells. Studies have shown that long‐term MR tracking of rapidly dividing cells underestimates their migration distance. Time‐lapse microscopy of random cellular motility and cell division was performed to evaluate the effects of SPIO‐labeling on neural stem cell migration. Labeled cells divided symmetrically and exhibited no changes in cell viability, proliferation, or apoptosis. However, SPIO‐labeling resulted in decreased motility of neural stem cells as compared with unlabeled controls. When SPIO‐labeled neural stem cells and human induced pluripotent stem cells were transplanted into mouse brain, rapid exocytosis of SPIO by live cells was observed as early as 48 h postengraftment, with SPIO‐depleted cells showing the farthest migration distance. As label dilution is negligible at this early time point, we conclude that MRI underestimation of cell migration can also occur as a result of reduced cell motility, which appears to be mitigated following SPIO exocytosis. Magn Reson Med, 2013. © 2012 Wiley Periodicals, Inc.     

Gadolinium-fullerenol as a paramagnetic contrast agent for cellular imaging

OBJECTIVES: The objectives of this study were to test cell-labeling methods to achieve intracellular labeling and T1 enhancement of cells on magnetic resonance imaging using a paramagnetic Gd@C82 fullerenol contrast agent, and to determine the effect of labeling on cell viability, metabolism, and differentiation capacity. MATERIALS AND METHODS: We tested the use of a transfection agent for labeling cells in culture with Gd@C82 fullerenol. Proliferation, viability, and differentiation assays of mesenchymal stem cell (MSC) cultures; light and electron microscopy of MSC and macrophages; and MRI of MSC, macrophage, and HeLa cervical carcinoma cell cultures in vitro and in vivo were performed to evaluate the labeled cells. RESULTS: Protamine sulfate transfection increased cell uptake of Gd@C82 fullerenols. The label was distributed in endosomes in the cytoplasm as shown by electron microscopy. High viability was shown for all cell lines and normal differentiation capacity was shown for MSCs. T1 of labeled MSC at 7 T was reduced 71% compared with unlabeled cells. CONCLUSIONS: Cellular labeling with Gd@C82 is feasible and can produce T1-enhanced cells on magnetic resonance imaging. This study suggests that further investigation of Gd fullerenols for tracking studies of viable cells, including stem cells, is warranted.     

Gradient echo acquisition for superparamagnetic particles with positive contrast (GRASP): sequence characterization in membrane and glass superparamagnetic iron oxide phantoms at 1.5T and 3T.

Iron oxides are used for cell trafficking and identification of macrophages in plaque using MRI. Due to the negative contrast, differentiation between signal loss caused by iron and native low signal in tissue may be problematic. It is, therefore, preferable to achieve positive contrast. The purpose of this study was to test the efficacy of a new MRI sequence GRASP (GRe Acquisition for Superparamagnetic Particles) to generate a positive signal in phantoms containing iron. Membrane phantoms were constructed containing Ferumoxide at 7 concentrations. Standard GRE sequences were modified with user controlled z-gradient rephasing (+/- 100%). CNR values were determined as a function of echo time (TE) and % rephasing at 1.5T and 3T. T(2)* values were determined using multiple double-echo GRE. The GRASP sequence generated positive signal enhancement in phantoms containing iron. For all rephasing values 相似文献   

心脏磁共振在体示踪移植细胞的可行性研究

目的：寻找一种能够对移植细胞进行在体示踪的标记方法,为移植细胞存留、迁移提供重要观察手段。方法：从中华小型猪髂骨处抽取骨髓,体外培养扩增骨髓间充质干细胞（MSCs）。将SPIO和MSCs共同孵育培养36h。普鲁士蓝染色评价细胞的标记效率;通过MTT比色实验评价SPIO对细胞生长能力的影响;台盼蓝染色检验标记后细胞的活性;使用Costar Transwell方法评价铁离子对细胞迁移能力的影响;用细胞分化诱导液培养标记后的细胞评价其向成脂肪细胞和成骨细胞的分化能力。在体内实验中将SPIO标记或未标记的自体MSCs注射到心肌内,通过心脏磁共振检查对移植细胞进行在体示踪观察,取材动物心脏行病理检查观察移植细胞的存活、存留。结果：MSCs经铁离子标记后普鲁士蓝染色阳性率在98%以上,可见蓝色颗粒位于细胞浆内,标记细胞电镜切片可见高密度铁颗粒位于细胞浆内。随着培养液中SPIO浓度的增加细胞增殖能力没有明显改变;标记后98%的细胞保持活性;SPIO标记后的细胞保持原有的形态,可继续培养、传代;SDF-1和VEGF诱导的迁移实验发现标记细胞迁移能力没有降低;铁离子标记后细胞仍可向成脂肪细胞和成骨细胞分化。注射到心肌内的SPIO标记的MSCs可通过心脏磁共振检查进行在体示踪,动态观察显示SPIO标记细胞在磁共振图像上表现为低信号,并且在移植后4周仍可成像。病理学检查可以看到移植细胞呈普鲁士蓝染色阳性,并和影像学有很好的一致性。结论：临床使用的SPIO磁共振对比剂可以安全、有效地标记MSCs,心脏磁共振检查可以实现SPIO标记的移植细胞的在体示踪。     

Relaxation effects of ferucarbotran‐labeled mesenchymal stem cells at 1.5T and 3T: Discrimination of viable from lysed cells

Human mesenchymal stem cells (hMSCs) were labeled with Ferucarbotran by simple incubation and cultured for up to 14 d. Iron content was determined by spectrometry and the intracellular localization of the contrast agent uptake was studied by electron and confocal microscopy. At various time points after labeling, ranging from 1 to 14 d, samples with viable or lysed labeled hMSCs, as well as nonlabeled controls, underwent MRI. Spin‐echo (SE) and gradient‐echo (GE) sequences with multiple TRs and TEs were used at 1.5T and 3T on a clinical scanner. Spectrometry showed an initial iron oxide uptake of 7.08 pg per cell. Microscopy studies revealed lysosomal compartmentalization. Contrast agent effects of hMSCs were persistent for up to 14 d after labeling. A marked difference in the T₂ effect of compartmentalized iron oxides compared to free iron oxides was found on T₂‐weighted sequences, but not on T‐weighted sequences. The observed differences may be explained by the loss of compartmentalization of iron oxide particles, the uniformity of distribution, and the subsequent increase in dephasing of protons on SE images. These results show that viable cells with compartmentalized iron oxides may—in principle—be distinguished from lysed cells or released iron oxides. Magn Reson Med, 2009. © 2009 Wiley‐Liss, Inc.     

Targeting EGFR-overexpressing tumor cells using Cetuximab-immunomicelles loaded with doxorubicin and superparamagnetic iron oxide

Epidermal growth factor receptor (EGFR), a cellular transmembrane receptor, plays a key role in cell proliferation and is linked to a poor prognosis in various human cancers. In this study, we constructed Cetuximab-immunomicelles in which the anti-EGFR monoclonal antibody was linked to poly(ethylene glycol)-block-poly(?-caprolactone) (PEG–PCL) nanomicelles that were loaded with doxorubicin (DOX) and superparamagnetic iron oxide (SPIO). The specific interactions between EGFR-overexpressing tumor cells (A431) and immunomicelles were observed using confocal laser scanning microscopy (CLSM) and flow cytometry. Furthermore, the capacity of transporting SPIO into tumor cells using these immunomicelles was evaluated with a 1.5 T clinical magnetic resonance imaging (MRI) scanner. It was found that the acquired MRI T2 signal intensity of A431 cells that were treated with the SPIO-loaded and antibody-functionalized micelles decreased significantly. Using the thiazolyl blue tetrazolium bromide (MTT) assay, we also demonstrated that the immunomicelles inhibited cell proliferation more effectively than their nontargeting counterparts. Our results suggest that Cetuximab-immunomicelles are a useful delivery vehicle for DOX and SPIO to EGFR-overexpressing tumor cells in vitro and that Cetuximab-immunomicelles can serve as a MRI-visible and targeted drug delivery agent for better tumor imaging and therapy.     

低浓度超顺磁性氧化铁标记兔骨髓间充质干细胞的体外磁共振成像研究

目的:以浓度为25μg Fe/ml的超顺磁性氧化铁纳米粒子(SPIO)体外标记兔骨髓间充质干细胞(BMSCs),并探讨1.5 T核磁共振仪成像的特征和成像所需最低标记细胞浓度,以及在标记后1 d、1周、2周、3周、4周的信号变化特征。方法:分离、纯化、培养兔BMSCs并以25μg Fe/ml的SPIO培养液浓度标记,对标记后不同时间的细胞行普鲁士蓝染色和台盼蓝拒染后显微镜观察,并进行MR成像,测量不同序列下不同浓度标记细胞管的信号强度,以确定扫描敏感序列及成像所需最低标记细胞浓度;再测量不同细胞浓度不同时相信号强度,来观察信号强度随时间变化的规律,并进行统计学分析。结果:浓度为25μg Fe/ml的超顺磁性氧化铁纳米粒子标记BMSCs的有效率接近100%,普鲁士蓝染色见细胞浆内有大小不等的蓝染铁颗粒,且在标记后4周内细胞仍具有活力,标记后的BMSCs在T2WI、尤其是GRE(T2*WI)序列信号明显降低;并且细胞浓度越高信号降低越明显,GRE序列MR成像的最低细胞浓度为5×104/ml。当标记细胞浓度为5×104/ml时,信号在T2*WI序列的降低2周后失去统计学意义;而在细胞浓度为5×105/ml时,标记3周后,信号在T2*WI序列的降低才失去统计学意义。结论:25μg/ml铁浓度标记干细胞不仅标记效率高,而且对细胞生长及增殖活力无明显影响,标记后MR信号改变与干细胞数目及标记时间相关。     

磁性标记的小胶质细胞在大鼠脑内的MR示踪

目的 探讨超顺磁性氧化铁颗粒(SPIO)标记的小胶质细胞在正常大鼠及阿尔茨海默病(AD)大鼠体内移植后,MR活体示踪的可行性.方法 以日本血液凝集病毒包膜(HVJ-E)为标记载体,将SPIO标记的小胶质细胞经颈内动脉注入正常大鼠(5只)及AD大鼠动物模型(5只)体内,3 d后应用7.0 T MR行T2*序列扫描,并与脑组织切片组织化学染色结果对照.结果 在正常大鼠脑内,MRI可见数个点状的信号改变区,这些信号改变区散在地分布在脑内各处,脑组织切片显示铁颗粒标记细胞位置与信号改变部位一致.MRI能够检测到由数个标记细胞引起的信号强度的改变.在AD大鼠模型脑内,MRI可见β-淀粉样蛋白42(Aβ42)注射区信号强度明显下降,信号改变区面积较大.与之相比,生理盐水注射区信号改变的强度及面积均不如Aβ42注射区改变明显.Aβ42注射区的标记细胞数为(454±47)个/mm2,明显高于生理盐水注射区的标记细胞数(83±13)个/mm2(P<0.05).结论 MRI可作为一种非侵入性检测手段在活的动物体内追踪标记细胞,在AD细胞水平的治疗中具有一定临床应用前景.     

Dual-Modal Nanoprobes for Imaging of Mesenchymal Stem Cell Transplant by MRI and Fluorescence Imaging

Objective

To determine the feasibility of labeling human mesenchymal stem cells (hMSCs) with bifunctional nanoparticles and assessing their potential as imaging probes in the monitoring of hMSC transplantation.

Materials and Methods

The T1 and T2 relaxivities of the nanoparticles (MNP@SiO₂[RITC]-PEG) were measured at 1.5T and 3T magnetic resonance scanner. Using hMSCs and the nanoparticles, labeling efficiency, toxicity, and proliferation were assessed. Confocal laser scanning microscopy and transmission electron microscopy were used to specify the intracellular localization of the endocytosed iron nanoparticles. We also observed in vitro and in vivo visualization of the labeled hMSCs with a 3T MR scanner and optical imaging.

Results

MNP@SiO₂(RITC)-PEG showed both superparamagnetic and fluorescent properties. The r₁ and r₂ relaxivity values of the MNP@SiO₂(RITC)-PEG were 0.33 and 398 mM^-1 s^-1 at 1.5T, respectively, and 0.29 and 453 mM^-1 s^-1 at 3T, respectively. The effective internalization of MNP@SiO₂(RITC)-PEG into hMSCs was observed by confocal laser scanning fluorescence microscopy. The transmission electron microscopy images showed that MNP@SiO₂(RITC)-PEG was internalized into the cells and mainly resided in the cytoplasm. The viability and proliferation of MNP@SiO₂(RITC)-PEG-labeled hMSCs were not significantly different from the control cells. MNP@SiO₂(RITC)-PEG-labeled hMSCs were observed in vitro and in vivo with optical and MR imaging.

Conclusion

MNP@SiO₂(RITC)-PEG can be a useful contrast agent for stem cell imaging, which is suitable for a bimodal detection by MRI and optical imaging.     

Magnetic resonance imaging detects differences in migration between primary and immortalized neural stem cells

RATIONALE AND OBJECTIVES: The study was performed to evaluate the effect of magnetic resonance imaging (MRI) contrast agent (super paramagnetic iron oxide [SPIO]) on differentiation and migration of primary murine neural stem cells (NSCs) in comparison to a neural stem cell line (C17.2). Because detection of labeled cells depends on the concentration of SPIO particles per imaging voxel, the study was performed at various concentrations of SPIO particles to determine the concentration that could be used for in vivo detection of small clusters of grafted cells. MATERIALS AND METHODS: Murine primary NSCs or C17.2 cells were labeled with different concentrations of SPIO particles (0, 25, 100, and 250 mug Fe/mL) and in vitro assays were performed to assess cell differentiation. In vivo MRI was performed 7 weeks after neonatal transplantation of labeled cells to evaluate the difference in migration capability of the two cell populations. RESULTS: Both the primary NSCs and the C17.2 cells differentiated to similar number of neurons (Map2ab-positive cells). Similar patterns of engraftment of C17.2 cells were seen in transplanted mice regardless of the SPIO concentration used. In vivo MRI detection of grafted primary and C17.2 cells was only possible when cells were incubated with 100 mug/mL or higher concentration of SPIO. Extensive migration of C17.2 cells throughout the brain was observed, whereas the migration of the primary NSCs was more restricted. CONCLUSIONS: Engraftment of primary NSCs can be detected noninvasively by in vivo MRI, and the presence of SPIO particles do not affect the viability, differentiation, or engraftment pattern of the donor cells.     

Superparamagnetic iron oxide-mediated hepatic signal intensity change in patients with and without cirrhosis: pulse sequence effects and Kupffer cell function

PURPOSE: To analyze superparamagnetic iron oxide (SPIO)-mediated hepatic signal intensity change in cirrhotic and noncirrhotic liver and to investigate the relationship between pulse sequence effects in SPIO-enhanced magnetic resonance (MR) imaging for hepatic cirrhosis. MATERIALS AND METHODS: Twelve patients with and 12 patients without cirrhosis underwent T2-weighted fast spin-echo, T2*-weighted gradient-echo (GRE), and T1-weighted GRE MR imaging before and twice (early and late phase) after SPIO administration. To assess the effect of SPIO, postcontrast relative signal-to-noise ratio (SNR) changes were statistically analyzed with repeated measurements analysis of variance for each pulse sequence. RESULTS: No interaction was shown between groups and data time points for any pulse sequence. There was no significant difference in mean hepatic relative SNR change on T2-weighted fast spin-echo images between the cirrhotic group and noncirrhotic group (-38.6% and -40.7%, early phase; -42.2% and -49.6%, late phase, respectively). For GRE images, statistically significant differences in mean hepatic relative SNR change were found between the cirrhotic group and noncirrhotic group (-14.2% and -44.5%, early phase; -28.5% and -56.4%, late phase on T2*-weighted GRE images (P <.001); 31.8% and 12.9%, early phase; 23.8% and 2.2%, late phase on T1-weighted GRE images (P <.05), respectively. CONCLUSION: Decreased overall phagocytic activity in cirrhotic liver is more likely due to Kupffer cell dysfunction than to Kupffer cell depletion, since magnetic susceptibility effects on T2*-weighted GRE images depend on intracellular SPIO cluster size.     

Staging of rectal cancer by double-contrast MR imaging using the rectally administered superparamagnetic iron oxide contrast agent ferristene and IV gadodiamide injection: results of a multicenter phase II trial

The aim of this study was to assess the accuracy of double-contrast magnetic resonance imaging (MRI) with rectal application of the superparamagnetic iron oxide contrast agent (SPIO) ferristene and IV gadodiamide for preoperative staging of rectal cancer. In a randomized phase II dose-ranging trial, 113 patients were studied preoperatively with one of four different formulations of ferristene (Abdoscan) as an enema before MRI. T1-weighted spin-echo (T1w SE) and T2w turbo spin-echo (TSE) single-contrast images were obtained as well as T1w SE and gradient-echo (GRE) double-contrast images after IV gadodiamide injection (Omniscan). Images were assessed qualitatively, and TNM tumor stage was compared with histopathology. High-viscosity ferristene formulations were superior to low-viscosity formulations in tumor staging (accuracy 90% vs 74%, P < 0.01). There was no significant difference between high and low iron content ferristene. MRI had a sensitivity of 97%, specificity of 50%, and accuracy of 82% for staging of rectal carcinoma higher than T2 stage. At receiver operator characteristic (ROC) analysis, MR differentiation between T1/T2 and T3/T4 tumor stages yielded a ROC index of 0.848. Double-contrast MRI is an accurate method for preoperative staging of rectal cancer.     

Comparison of SPIO and USPIO for in vitro labeling of human monocytes: MR detection and cell function

PURPOSE: To label human monocytes with superparamagnetic iron oxide (SPIO) and compare labeling efficiency with that of ultrasmall SPIO (USPIO) and evaluate the effect of iron incorporation on cell viability, migratory capacity, and proinflammatory cytokine production. MATERIALS AND METHODS: The study was approved by the institutional ethics committee; informed consent was obtained from donors. Freshly isolated human monocytes were labeled with iron particles of two sizes, USPIOs of 30 nm and SPIOs of 150 nm, for 1.5 hours in culture medium containing 0.1, 0.5, 1.0, and 3.7 mg of iron per milliliter. Labeling efficiency was determined with relaxation time magnetic resonance (MR) imaging (4.7 T) and Prussian blue staining for presence of intracellular iron. Cell viability was monitored; migratory capacity of monocytes after labeling was evaluated by using an in vitro assay with monolayers of brain endothelial cells. Levels of proinflammatory cytokines, interleukin (IL) 1 and IL-6, were measured with enzyme-linked immunosorbent assay 24 hours after labeling. Data were analyzed with Student t test or two-way analysis of variance followed by a multiple-comparison procedure. RESULTS: R2 relaxation rates increased for cell samples incubated with SPIOs, whereas rates were not affected for samples incubated with highest concentration of USPIOs. Labeling monocytes with SPIOs (1.0 mg Fe/mL) resulted in an R2 of 13.1 sec(-1) +/- 0.8 (standard error of the mean) (7 sec(-1) +/- 0.2 for vehicle-treated cells, P < .05) and had no effect on cell viability. On the basis of T2 relaxation times, the in vitro MR detection limit of 58 labeled monocytes per 0.05 microL was calculated. Migration of labeled monocytes was not different from that of vehicle-treated cells. Intracellular iron had no effect on production of IL-1 and IL-6 24 hours after labeling. CONCLUSION: In vitro labeling of human monocytes is effective by using SPIOs, not USPIOs. Incubation with SPIOs (1.0 mg Fe/mL) results in efficient labeling detectable on MR images and does not affect cellular viability and activation markers such as cell migration and cytokine production.     

Hepatic and hepatocarcinoma magnetic resonance: comparison of the results obtained with paramagnetic (gadolinium) and superparamagnetic (iron oxide particles) contrast media

PURPOSE: To compare prospectively dynamic gadolinium (Gd)-enhanced with superparamagnetic iron oxide (SPIO)-enhanced MRI for the detection of hepatocellular carcinoma (HCC). MATERIAL AND METHODS: Twenty-five patients with histologically proven HCC and liver cirrhosis (28% of them in B or C Child class) underwent dynamic Gd-enhanced MRI and, a few days later, (mean interval: three days) SPIO-enhanced MRI. Only patients with availability of clinical and imaging follow-up for at least seven months were enrolled in this prospective study. Axial dynamic Gd-enhanced imaging was performed with T1 gradient-recalled echo (GRE) sequences. Both axial and sagittal SPIO-enhanced imaging were performed with respiratory triggered T2-weighted turbo spin-echo (TSE) and T1-T2*-weighted GRE sequences. MR images were reviewed by two independent radiologists. The readers scored each lesion for the presence of HCC and assigned confidence levels based on a five-grade scale: 1, definitely or almost definitely absent; 2, possibly present; 3, probably present; 4, definitely present; 5, definitely present with optimal liver/lesion contrast or good liver/lesion contrast and morphological signs (intact capsule, intranodular septa, extra-capsular infiltration), useful for locoregional treatment planning. A positive diagnostic value was assessed for scores of 3 or higher. RESULTS: Gd-enhanced and SPIO-enhanced MRI found 44 lesions. The combined use of TSE and GRE SPIO-enhanced sequences detected 11 more lesions (25% improvement in sensitivity) than Gd-enhanced MRI. One lesion (2.27%) was detected only with Gd-enhanced MRI. Eight of twelve lesions visible with a single contrast agent measured less than 1 cm in diameter. HCC detectability was 75% with Gd-enhanced MRI and 97.7% with SPIO-enhanced MRI. SPIO-enhanced T2-weighted TSE images showed significantly higher diagnostic value than SPIO-enhanced T1-T2*GRE images only in three cases, while nodule morphological characteristics (capsule, septa, different cell differentiation components) were better depicted by TSE images. DISCUSSION AND CONCLUSIONS: In our study the combined use of SPIO-enhanced T2-weighted TSE and T1-T2*-weighted GRE sequences showed higher sensitivity than gadolinium-enhanced GRE dynamic imaging (97.7% versus 75%). These results are at least partly related to our study conditions, that is: 1) MRI was performed with a 1T system, 2) both axial and sagittal SPIO-enhanced imaging were performed with respiratory triggered T2-weighted TSE and T1-T2*-weighted GRE, 3) there was a low freaquency of severe cirrhosis.     

In vitro comparison of the biological effects of three transfection methods for magnetically labeling mouse embryonic stem cells with ferumoxides.

In vivo MRI of stem cells (SCs) is an emerging application to evaluate the role of cell therapy in restoring the injured myocardium. The high spatial and temporal resolution combined with iron-oxide-based intracellular labeling techniques will provide a sensitive, noninvasive, dual imaging modality for both cells and myocardium. In order to facilitate this novel imaging approach, much effort has been directed towards developing efficient transfection methods. While techniques utilizing poly-L-lysine (PLL), protamine sulfate (PS), and electroporation (ELP) have been proposed, the fundamental biological effects of these methods on mouse embryonic SCs (mESC) have not been investigated systematically. In this study a longitudinal in vitro evaluation of cellular viability, apoptosis, proliferation, and cardiac differentiation of magnetically labeled mESC was conducted. No significant difference was seen in these biological parameters among the three transfection methods. However, cardiac differentiation was most attenuated by ELP, and iron uptake was most effective by PS.